Abstract
[3H]-labelled Casein and chromogenic substrates were used for a quantitative determination of protease activity in cell culture supernatants during long-term cultivation of BHK and hybridoma cells. They produced recombinant human interleukin 2 and an IgG2a - antibody respectively in serum-or protein free medium. Characterization of protease activities was performed by inhibitor studies and specific p-NA derivates. Only 20% of the total protease activity in hybridoma and up to 50% in BHK cell supernatants is based on serine type proteases. Supplementation of supernatants with 2-mercaptoethanol or magnesiumchloride caused an increase of protease activities.
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References
Conradt, H. S.; Ausmeier, M.; Dittmar, K. E.; Hauser, H. J.; Lindenmaier, W. (1986) Secretion of Glycosylated Human Interleukin-2 by Rekombinant Mammalian Cell Lines, Carbohydrate Research 149, pp 443–450
Goldberg, A. R. (1974) Increased Protease Levels in Transformed Cells: A Casein Overlay Assay for the Detection of Plasminogen Activator Production, Cell 2, pp 95–102
Hatcher, V. B.; Wertheim, M. S.; Rhee, C. Y.; Tsien, G.; Burk, P. G. (1976) Relationship between Cell Surface Protease Activity and Doubling Time in Various Normal and Transformed Cells, Biochim. et Biophys. Acta 451, pp 499–510
Hatcher, V. B.; Obermann, M. S.; Wertheim, M. S.; Rhee, C. Y.; Tsien, G.; Burk, P. G. (1977) The Relationship between Surface Protease Activity and the Rate of Cell Proliferation in Normal and Transformed Cells, Biochem. and Biophys. Res. Comm. 76, pp 602–608
Jäger, V. (1988) Entwicklung eines Hohlfaser-Perfusionsreaktor-Systems zur Produktionsoptimierung monoklonaler Antikörper, Dissertation, Universität Hannover
Kás, J., Rauch, P. (1982) Radioisotopic Method for the Determination of Low and Very Low Proteolytic Activities in Foodstuffs. Z. Lbensm. Unters. Forsch. 174, 290–293
Kassel, B.; Wang, T.-W. (1970) The Action of Thermolysin on the Basic Trypsin Inhibitor of Bovine Organs; in: Proc. Int. Res. Conf. of Proteinase Inhibitors, München, pp. 89–94, WdG Verlag, Berlin, 1971
Lehmann, J.; Vorlop J.; Büntemeyer H. (1988) Bubble-free Reactors and their Development for Continuous Culture with Cell Recycle. In: Animal Cell Biotechnology (R.E. Spier and J.B. Griffith eds.) Vol. 3, pp 221–237
Lucid-Lange, M.; Wagner, R. (1990) Conditions for the Production of Recombinant IL-2 in Stirred Suspension Culture Using a Protein-Free Medium. In: Production of Biologicals from Animal Cells in Culture Research, Development and Achievements (Spier, R. E.; Griffiths, J. B.; eds.) Butterworths, in Press
Murakami, H. (1989) Serum-Free Media Used for Cultivition of Hybridomas. In Monoclonal Antibodies: Production and Application, pp 107–141; Alan. Liss Inc.
Neurath, H. (1984) Evolution of Proteolytic Enzymes, Science 224, pp 350–357
Ryll, T.; Lucki-lange, M.; Jäger, V.; Wagner, R. (1990) Production of Recombinant Interleukin 2 with BHK Cells in a Hollow Fibre and a Stirred Tank Reactor with Protein-Free Medium, J. Biotechnology 14, pp 377–392
Schlaeger, E. J.; Eggimann, B.; Gast, A. (1987) Proteolytic Activity in the Culture Supernatants of Mouse Hybridoma Cells, Develop. Biol. Standard 66, pp 403–408
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© 1991 Springer Science+Business Media Dordrecht
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Lind, W., Lietz, M., Jäger, V., Wagner, R. (1991). Proteolytic Activities in Serum-Free Supernatants of Mammalian Cell Lines. In: Sasaki, R., Ikura, K. (eds) Animal Cell Culture and Production of Biologicals. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-3550-4_37
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DOI: https://doi.org/10.1007/978-94-011-3550-4_37
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